Imobilization of scFv through SiN-binding peptide for sensitive RIfS sensor

• Project/Area Number: 26420795
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Kyoto Institute of Technology
• Principal Investigator: Kumada Yoichi 京都工芸繊維大学, 分子化学系, 准教授 (70452373)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: SiN / SiN-tag / antibody / GST / immobilization / orientation control / 窒化ケイ素 / 単鎖抗体 / 材料親和性ペプチド / 配向制御 / リフォールディング / 固定化 / バイオセンサ

Outline of Final Research Achievements

In this study, we developed a new immobilization method of protein onto the surface of silicone nitride. First, we successfully identified SIN-binding peptides from Elongation Factor Tu which was isolated from E. coli cell lysate. Especially, one of them showed the strongest binding affintiy towards the surface of SIN plate. By genetic fusion of SIN-tag to the C-termimus of single-chain Fv antibodies resulted in higher antigen-binding activity of scFv in adsorption state. Thus, the SIN-binding peptides developed in this study will be significantly useful for site-specific immobilization of recombinant antibody fragments such as scFvs in RIfS sensor.

Research Products

• [Journal Article] Identification and characterization of polydimethylsiloxane-binding peptides (PDMS-tag) for oriented immobilization of functional protein on a PDMS surface. (2016)
  • Author(s): Y. Kumada, R. Otsuki, Y. Sakoda, R. Akai, K. Matoba, J. Katayama, M. Kishimoto, J. Horiuchi,
  • Journal Title: J. Biotechnol. Volume: 236 Pages: 193-198
  • DOI: 10.1016/j.jbiotec.2016.08.005
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] 1.Efficient preparation and site-directed immobilization of VHH antibodies by genetic fusion of poly(methylmethacrylate)-binding peptide (PMMA-tag). (2015)
  • Author(s): Kumada Y, Kang B, Yamakawa K, Kishimoto M, Horiuchi JI
  • Journal Title: Biotechnology Progress Volume: 31 Issue: 6 Pages: 1563-1570
  • DOI: 10.1002/btpr.2169
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 組換え大腸菌を用いた活性型一本鎖抗体の高効率分泌生産 (2015)
  • Author(s): 李歓・岸本通雅・堀内淳一・熊田陽一
  • Organizer: 化学工学会 秋季大会
  • Place of Presentation: 北海道・札幌市
  • Year and Date: 2015-09-09